用DNA限制性片段长度多态性鉴定中国旋毛虫3个分离株

以旋毛虫国际标准虫株Ｔ．ｓｐｉｒａｌｉｓ（Ｔｓ）、Ｔ．ｎａｔｉｖａ（Ｔｎ）和Ｔ．ｎｅｌｓｏｎｉ（Ｔｎｅ）作对照，利用限制性片段长度多态性（ＲＦＬＰ）差异对分离自我国黑龙江省３株旋毛虫进行虫种归属鉴定。５种内切酶实验结果显示：黑龙江猪株（ＨＬ）与Ｔｓ酶切图谱相同；犬株（ＷＣ）与Ｔｎ酶谱相同；猫株（ＳＷ）与Ｔｎ的ＤｒａⅠ，ＰｓｔⅠ，ＨａｅⅢ酶谱相近，而与Ｔｎｅ酶谱差异显著。提示：黑龙江猪株系Ｔｓ；犬株系Ｔｎ；猫株在分类归属上似乎与Ｔｎ靠近。     

我国7株旋毛虫基因组DNA的研究

对我国７株旋毛虫基因组ＤＮＡ，采用５种限制性内切酶分析表明，来自猪体的旋毛虫酶谱相似，来自犬、猫体的旋毛虫酶谱相似，而猪体与犬、猫体的旋毛虫酶谱差异明显。同时采用随意扩增的多态ＤＮＡ分析，引物为：５＇ＡＡＴＴＣＧＣＧＧＣＣＧＣＧＴＣＧＡＣ３＇，各分离株显示自己特有带型。比较不同地区猪体或犬、猫体的旋毛虫及同一地区不同宿主的旋毛虫的相似率。提示：地区差异愈大，相似率愈小；宿主亲缘愈远，其相似率愈小。似乎我国旋毛虫可分为两群：一群是猪体旋毛虫，另一群是犬、猫体旋毛虫。     

中国大陆4地日本血吸虫蛋白质及同工酶的多态性研究

本文通过对中国大陆境内安徽、湖南、湖北和云南４地日本血吸虫的蛋白质组分、同工酶多态性进行分析表明：１梯度ＳＤＳ、ＩＥＦ电泳结果均显示４地日本血吸虫雌雄虫的蛋白条带相同，说明在蛋白质组分上可能是相同的。２同工酶电泳显示４地血吸虫雌雄虫在ＣＯ酶谱上无差别，即没有多态性表现。在ＧＰＩ、ＡＣＰ和Ｇ６ＰＤＨ上显示了雌雄虫间的酶谱略有差异，但不同地理株同性虫体间未显示差异，说明在雌雄虫间可能存在基因位点的多态性，在ＰＧＭ和ＭＤＨ的同工酶谱上则表现出既有雌雄虫间的不同，也有不同地理株间的差异，表现４地日本血吸虫雌雄虫和不同地理株间有基因位点的多态性表现。     

SOLUBLE PROTEIN AND ISOENZYME ANALYSIS OF FOUR GIARDIA ISOLATES FROM CHINA AND AMERICA

对四株蓝氏贾第虫［三株分离自中国（Ｃ１、Ｃ２、Ｃ３）］,一株分离自美国（ＣＤＣ）］的可溶性蛋白和同工酶分别用ＳＤＳ—ＰＡＧＥ和ＰＡＧＥ法进行分析。结果表明,每一受试虫株在ＳＤＳ—ＰＡＧＥ中均显示出众多蛋白带,其主带数目分别为９、１３、１３、１１。大多数带的分子量介于１７．５ＫＤａ～９４ＫＤａ之间。同工酶分析结果表明,在酯酶（ＥＳＴ）分析中,ＣＤＣ出现４条带,其中３条深染。Ｃ３呈现３条浅带,但Ｃ１和Ｃ２未见条带出现。在酸性磷酸酶（ＡＰ）分析中,三个中国虫株均各出现１条浅带和１条深带,而在ＣＤＣ中只出现１条。在苹果酸脱氢酶（ＭＤＨ）分析中,Ｃ１可见１条带,而在Ｃ３为２条,Ｃ２和ＣＤＣ未见条带。在苹果酸酶（ＭＥ）分析中,中国三个虫株均各出现１条带,而ＣＤＣ则无     

大鼠骶髓后连合核神经元兴奋性氨基酸诱导的反应

应用制霉菌素穿孔全细胞电压钳技术，研究急性分离的大鼠骶髓后连合核（ＳＤＣＮ）神经元对谷氨酸受体激动剂的反应。钳制电压为－４０ｍＶ的条件下，Ｌ－谷氨酸（Ｇｌｕ），Ｎ－甲基－Ｄ－门冬氨酸（ＮＭＤＡ），使君子酸（ＱＡ），α－氨基－３－羟基－５－甲基异倾阶－４－丙酸（ＡＭＰＡ）和红藻氨酸（ＫＡ）均诱导产生内向电流。随着激动剂浓度的增高，这些电流的量效关系曲线呈典型的Ｓ型。ＥＣ５０值分别是Ｇｌｕ３．３×１０－５Ｍ，ＮＭＤＡ９．０×１０－ＳＭ，ＱＡ６．４×１０７Ｍ，ＡＭＰＡ１．３×１０－４及ＫＡｇ．６×ｕ１０－５Ｍ。Ｎｉｌｌ系数分别是Ｇｌｕ０．７４，ＮＭＤＡ０．８３，ＱＡ１．３，ＡＭＰＡ１．１和ＫＡ１．３。代谢型谷氨酸受体激动剂ｔＡＣＰＤ（１０－３Ｍ）未能诱导出电流．Ｃｙｃｌｏｔｈｉａｚｉｄｅ显著增强ＫＡ和ＡＭＰＡ诱导的反应。相反，伴刀豆球蛋白Ａ（ＣｏｎＡ）对ＫＡ和ＡＭＰＡ反应作用甚微，提示ＳＤＣＮ神经元主要表达ＡＭＰＡ型非ＮＭＤＡ受体。     

天津地区G6PD缺陷患者常见基因突变分析

目的研究北方地区葡萄糖－６－磷酸脱氢酶（ｇｌｕｃｏｓｅ－６－ｐｈｏｓｐｈａｔｅｄｅｈｙｄｒｏｇｅｎａｓｅ，Ｇ６ＰＤ）基因突变类型、基因突变与人口变迁。方法采用“错配引物”介导的聚合酶链反应／限制性酶切分析法和双脱氧核苷酸指纹印迹检测法，对天津地区２２例Ｇ６ＰＤ缺陷患者进行３种中国南方人群常见的Ｇ６ＰＤ基因突变的分析。结果８例患者（８／２２，３６．４％）有Ｒ４５９Ｌ（１３７６Ｇ→Ｔ）突变；７例患者（７／２２，３１．８％）有Ｒ４６３Ｈ（１３８８Ｇ→Ａ）突变；７例（７／２２）无１３７６和１３８８位点突变病例中，有３例做了Ｈ３２Ｒ（９５Ａ→Ｇ）突变分析，提示１例有该位点突变。结论北方地区也存在中国南方人群常见的３种Ｇ６ＰＤ基因突变，大多数中国北方Ｇ６ＰＤ缺陷患者是由于中国南方移民迁移造成的。     

急性丙型肝炎患者免疫状况的研究

用间接免疫荧光法、酶联免疫吸附试验（ＥＬＩＳＡ）及ＬＤＨ释法，对１６例急性输血后丙型肝炎（抗－ＨＣＶ、ＨＣＶ－ＲＮＡ均阳性）患者，分别进行外周血单个核细胞（ＰＢＭＣ）的Ｔ细胞亚群计数、Ｔ４／Ｔ８比值、Ｔａｃ受体的检测及血清可溶性白细胞介素２受体（ｓＩＬ－２Ｒ）ＮＫ细胞活性的测定。并与正常人组比较，经ｔ检验发现，急性丙型肝炎患者的Ｔ４亚群所占百分比、Ｔ４／Ｔ８比值及ＰＢＭＣＴａｃ受体表达均明显低于正常人组（Ｐ＜０．０５），而ＮＫ细胞活性、血清ｓＩＬ－２Ｒ明显高于正常人组（Ｐ＜０．０５）。患者的这些免疫状态改变，可能对其发病机理的研究有一定意义。     

一株与正常人细胞成分反应的巨细胞病毒单克隆抗体的鉴定

用ＨＣＭＶ－ＡＤ１６９株抗原建立了一株分泌ＨＣＭＶＭｃＡｂ的杂交瘤细胞株Ｎ４Ｅ６。经Ｗｅｓｔｅｒｎｂｌｏｔ分析发现ＭｃＡｂ与ＨＣＭＶ抗原和正常人成纤维细胞（ＨＥＦ）抗原均有多条免疫沉淀带出现：与ＨＣＭＶ－ＡＤ１６９株抗原反应的沉淀带为８９Ｋ，８７Ｋ，７３Ｋ和４２Ｋ；与Ｄａｖｉｓ株抗原反应的为８９Ｋ，８７Ｋ，４４Ｋ；与８１６地方株抗原反应的为８９Ｋ，８７Ｋ，７３Ｋ，４４Ｋ；与ＨＥＦ抗原反应的为８９Ｋ，８７Ｋ，４４Ｋ。表明Ｎ４Ｅ６除能识别ＨＣＭＶ－ＡＤ１６９株和８１６地方株特异成分７３Ｋ，外，尚能识别ＨＣＭＶ和ＨＥＦ共同成分８９Ｋ，８７Ｋ和４４Ｋ。     

我国旋毛虫Ts254 DNA探针制备及序列测定

我国云南猪体旋毛虫分离株（ＢＳ）经随意扩增的多态ＤＮＡ（ＲＡＰＤ）方法产生的２５４ｂｐ条带，以地高辛（ｄｉｇｏｘ－ｉｎｉｎ）标记制成探针（Ｔｓ２５４），将此探针与７株旋毛虫总ＤＮＡ进行斑点杂交，出现明显的杂交斑点，检出的ＤＮＡ量为１ｎｇ。但与其它相关寄生虫、质粒ｐＢＲ３２２、昆明株小鼠肌肉ＤＮＡ均未出现杂交反应，表明Ｔｓ２５４探针具有一定的特异性和较高的敏感性。将此Ｔｓ２５４ＤＮＡ片段克隆进Ｍ１３ｍｐ１８／１９噬菌体，扩增后，筛选阳性重组子，抽提单链后测序。经Ｇｅｎ－Ｂａｎｋ序列数据库检索，Ｔｓ２５４序列与真核ＤＮＡ及结构ＲＮＡ序列无有意义的同源性，此旋毛虫ＤＮＡ序列为我国首次报道。     

他巴唑对Graves‘病患者血清可溶性gp130的影响

５２例Ｇｒａｖｅｓ’病（ＧＤ）患者随机分为两组，２６例接受他巴唑（ＭＭＩ）治疗，２６例接受ＭＭＩ加心得安治疗。应用酶联免疫吸附法（ＥＬＩＳＡ），测定了未治疗（第０周）、部分缓解（第２周）和缓解组（第６周）患者及２０例健康对照者血清可溶性ｇｐ１３０（ｓｇｐ１３０）水平。结果表明：①ＭＭＩ和ＭＭＩ加心得安治疗的未治疗组、部分缓解组患者血清ｓｇｐ１３０水平显著高于健康对照组（Ｐ＜０．０１），缓解组与健康对照者血清ｓｇｐ１３０水平无显著差异（Ｐ＞０．０５）；②ＭＭＩ与ＭＭＩ加心得安治疗患者对应组的血清ｓｇｐ１３０水平无差异（分别Ｐ＞０．０５）；③ＧＤ患者血清ｓｇｐ１３０与游离Ｔ３、游离Ｔ４水平呈正相关（分别ｒ＝０．３１８，ｒ＝０．２９７，Ｐ＜０．０５）。提示：ＭＭＩ对ＧＤ患者血清ｓｇｐ１３０的产生具有抑制作用     

Morphologic, biometric, and isoenzyme characterization of Trichuris suis

Trichuris suis isolates were collected from the cecum of Sus scrofa domestica (pig) and S. s. scrofa (wild boar). Morphology and biometry studies were carried out. Morphology studies showed the existence of typical caudal papillae in males of T. suis from wild boars, but no other difference was observed in the biometric parameters (total length, esophageal length, posterior-portion body length, and spicular length) of T. suis isolated from either host. Individual extracts were subjected to malate dehydrogenase (MDH), malic enzyme (ME), glucose 6-phosphate dehydrogenase (G6PD), lactate dehydrogenase (LDH), and superoxide dismutase (SOD) isoenzyme analysis following starch-gel electrophoresis, and the isoenzyme patterns were compared with those obtained from other species of trichurids. MDH, ME, G6PD, LDH, and SOD isoenzyme patterns were identical for T. suis from both hosts. MDH isoenzyme patterns were characterized by the presence of one cathodic isoenzyme. ME, G6PD, and LDH isoenzyme patterns indicated the presence of three phenotypes, whereas the SOD isoenzyme pattern showed only one phenotype characterized by the existence of two (anodic and cathodic) bands. Different LDH and SOD isoenzyme patterns observed for T. suis, T. ovis, and T. skrjabini confirm once more that isoenzyme patterns have potential as a diagnostic tool for differentiation of different species of Trichuris. Received: 21 October 1997 / Accepted: 2 December 1997     

Morphological and genetic differentiation of Rodentolepis straminea (Goeze, 1752) and Rodentolepis microstoma (Dujardin, 1845) (Hymenolepididae)

The two related species, Rodentolepis straminea (Goeze, 1782) and Rodentolepis microstoma (Dujardin, 1845) (Cestoda, Hymenolepididae), both parasites of rodents, were compared morphologically and electrophoretically. Adult worms were isolated from three wild rodent species of the family Muridae (Apodemus flavicollis, Apodemus sylvaticus, and Mus musculus) from three different sites in Spain and France. Although these two species were strikingly similar in morphological appearance, some of the morphological and metrical features analysed (scolex, mature segments and eggs) can be used for differentiation. Fixed allelic differences were found. Of the ten enzymes detected by starch-gel electrophoresis, six (AAT, AK, GPI, MDH, NP, PGM) showed characteristic isoenzyme profiles in each species. Only in MPI, PEPC, PEPD, and ME enzyme loci were no differences found. The study revealed that the two taxa can be clearly differentiated.     

Isoenzyme variation within the genusCryptosporidium

Soluble extracts of the oocysts ofCryptosporidium parvum had demonstrable, but low, activities of malate dehydrogenase (MDH, EC. 1.1.1.37), carboxylesterase (ES, EC 3.1.1.1) and lactate dehydrogenase (LDH, EC. 1.1.1.27) following thin-layer starch-gel electrophoresis. Much higher activities of glucose phosphate isomerase (GPI, EC. 5.3.1.9) and phosphoglucomutase (PGM, EC. 2.7.5.1) were found, and zymograms of these two enzymes were used to characterise isolates ofC. parvum from human, bovine, ovine and cervine sources,C. muris from the brown rat andC. baileyi from young turkeys. PGM and GPI zymograms clearly distinguished betweenC. parvum, C. muris andC. baileyi. The five isolates ofC. parvum showed the same electrophoretic mobility for GPI, whereas the PGM mobilitiy of the single human isolate ofC. parvum examined was clearly different from that of the other isolates. This is the first report of the use of isoenzymes to distinguish between species and isolates ofCryptosporidium.     

Hybrids betweenSchistosoma mansoni andS. rodhaini: characterization by isoelectric focusing of six enzymes

Results are reported of enzyme analyses by isoelectric focusing in polyacrylamide gels of individual extracts fromSchistosoma mansoni (Guadeloupe),S. rodhaini (Burundi), and their experimental hybrids (first and second generation). The distinctive patterns of lactate dehydrogenase (LDH), malate dehydrogenase (MDH), glucose-6-phosphate dehydrogenase (G6PD), acid phosphatase (AcP), phosphoglucomutase (PGM) and glucose phosphate isomerase (GPI) enable the characterization of the two parental strains and the hybrids. Particular observations, such as the existence of a polymorphism at both MDH-1 and MDH-2 loci and a sex-linked heredity for GPI, are discussed. A genetic interpretation is proposed to explain the patterns observed for MDH and GPI (with a dimeric structure) and for PGM (monomeric structure); a comparison is made with electrophoretic data available forS. mansoni andS. rodhaini.     

Leishmania (Viannia) naiffi sp. n., a parasite of the armadillo, Dasypus novemcinctus (L.) in Amazonian Brazil

A new leishmanial parasite, Leishmania (Viannia) naiffi sp. n., is described from the nine-banded armadillo, Dasypus novemcinctus (Edentata: Dasypodidae), from Para State, north Brazil. The parasite grows luxuriantly in Diffco blood-agar medium (B47), but poorly in the skin of intradermally inoculated hamsters. A comparison of isoenzyme profiles by starch gel electrophoresis separates the parasite from L. (V) braziliensis and L. (V.) guyanensis by the enzymes ASAT, ALAT, PGM, GPI, G6PD, PEP, MPI and GD, and from Leishmania (Leishmania) chagasi, L. (L.) amazonensis and L. (L.) deanei by ASAT, ALAT, PGM, GPI, MPI, G6PD, MDH, PEP and ACON. Finally, L. (V.) naiffi is serologically differentiated from L. (V.) braziliensis, L. (V.) guyanensis and L. (V.) panamensis on monoclonal antibodies specific for these parasites.     

Isoenzyme studies on cercariae from monoinfections and adult worms ofSchistosoma mansoni (10 isolates) andS. rodhaini (one isolate) by horizontal polyacrylamide gel electrophoresis and staining of eight enzymes

The enzymes phosphoglucose isomerase (PGI), phosphoglucomutase (PGM), hexokinase (HK), adenylate kinase (AK), fructokinase (FK), mannose-6-phosphate isomerase (MPI), glucose-6-phosphate dehydrogenase (G-6-PDH) and malate dehydrogenase (MDH) were chosen to study the variation between isolates, cercariae and adults, individuals, and sexes ofSchistosoma mansoni andS. rodhaini, using horizontal polyacrylamide gel electrophoresis. The method described allows combinations of six of the eight enzymes to be scored in the homogenate from one adult worm.In adultS. mansoni one phenotype of the eight enzymes was observed in all isolates. In addition, the enzyme PGI showed polymorphism in the isolates from Tala, Kenya and Uganda. PGM in the isolates from Tala, Kenya and South Africa showed polymorphism. The cercarial phenotype differs from the adult phenotype in G-6-PDH, where the cercarial enzyme mobility is slower than that in the adult worm. The low amount of intrastrain variation observed in this species is explained by the limited amount of material used to establish the laboratory stocks, whereas the genetic similarity between geographically widely separated stocks does suggest that only limited geographical variation is likely to occur inS. mansoni.It is suggested that the gene controlling the PGI polymorphism is located on the sex chromosomes ofS. mansoni.Mobility differences were observed betweenS. mansoni andS. rodhaini in the enzymes PGI and PGM, and these characteristics might be useful for a quick identification of schistosome cercariae emerging fromBiomphalaria sp. in Africa.     

Isoenzymatic characterization of Phlebotomus papatasi (Diptera: Psychodidae) of the Marrakech area, Morocco

This study reports the genetic characterization of urban and rural populations of Phlebotomus (Phlebotomus) papatasi Scopoli (Diptera: Psychodidae) in Marrakech, Morocco. Using isoenzymatic analysis, four Moroccan populations were compared with other Mediterranean basin populations from Spain, Cyprus, and Syria. Morphological anomalies were noted in the male genitalia of 5.3% of the specimens collected from Marrakech area. Qualitative analysis of zymogram profiles revealed nine polymorphic enzymes (HK, PGM, PGI, 6PGD, MDH1, MDH2, ICD2, FUM, and ACO) and three monomorphic enzymes (ME, ICD1, and alphaGPDH). Genetic distances clearly separated the populations of western Mediterranean countries (Morocco and Spain) from eastern countries (Syria and Cyprus), but they could not be used to differentiate between urban and rural populations in Marrakech area.     

Observations on an isolate ofSchistosoma bovis from Tanzania

The eggs ofSchistosoma bovis isolated from Misungwi, Tanzania measure 211.1 m±18.4 long and 66.7 m±5.4 wide. The parasite is naturally transmitted byBulinus africanus and is compatible in the laboratory with snails belonging to theB. truncatus, B. forskali, andB. reticulatus groups. The compatibility withB. africanus group snails is shared with isolates from Kenya and Sudan but not withS. bovis from more northern distributions. Enzyme analyses were carried out by isoelectric focusing. In adult worms, phosphoglucomutase (PGM), hexokinase (HK), malate dehydrogenase (MDH), and lactate dehydrogenase (LDH) proved to be monomorphic whereas two types of glucosephosphate isomerase (GPI), three types of glucose-6-phosphate dehydrogenase (G6PDH), and two types of acid phosphatase (AcP) were identified. Differences in the pI values of GPI and MDH of snail digestive glands and of larval parasites allowed the intramolluscan stages to be characterised. The GPI heterogeneity encountered was common both to the larval and adult parasites. The enzyme types identified inS. bovis are discussed both from an intra- and interspecific viewpoint.     

Isolation and characterization of flagellates from rodents and canids in Masinga,Machakos District,Kenya

A total of 728 animals comprising of 633 rodents and 95 canids were examined for leishmanial parasites. Flagellates were isolated from 67 out of 111 (60.4%) Acomys subspinosus (spiny mouse), 12 out of 143 (8.4% ) Mastomys natalensis (multimammate rat), 2 out of 50 (4.0%) Lemniscomys striatus (striped mouse), 2 out of 6 (33.3%) Herpestes sanguineus (slender mongoose), 1 of 1 Helogale parvula (dwarf mongoose) and 1 out of 84 Canis familiaris (domestic dog). All isolates were characterized by Isoenzyme analysis using nine enzymes, namely, malate dehydrogenase (MDH), phosphoglucomutase (PGM), glucose phosphate isomerase (GPI), isocitrate dehydrogenase (ICD), nucleoside hydrolase (NH), glucose 6-phosphate dehydrogenase (G6PD), malic enzyme (ME), 6-phosphogluconate dehydrogenase (GPGD) and mannose phosphate isomerase (MPI). Enzyme profiles of these isolates were compared with those of five WHO Leishmania reference strains and five well characterized rodent trypanosomes of the subgenus Herpetosoma. The profiles of the isolates were found to be different from those of the Leishmania and Trypanosoma reference strains but the parasites were morphologically similar to rodent trypanosomes. These results suggest that Leishmania parasites were not among the isolates. The enzymes profiles of the three mongoose isolates were identical but differed from profiles of isolates from rodents and dog. This is the first time in Kenya that a high prevalence of nonpathogenic trypanosomes is reported in rodents and canids. From the epidemiological point of view, these trypanosomes must be differentiated from the pathogenic species of trypanosomes and Leishmania that infect man and other animals. The results of this study suggest that rodents do not seem to play a role as reservoirs of Leishmania parasites in Masinga Location, Kenya.     

Application of multilocus enzyme gel electrophoresis to Haemophilus influenzae.

Multilocus enzyme electrophoresis was adapted to the study of Haemophilus influenzae. Protein extracts from sonicated whole bacteria were subjected to starch gel electrophoresis. After staining with substrates, the position of each isoenzyme (electromorph) was registered. Each isolate was assigned an electrophoretic type (ET) by the combination of electromorphs for the enzymes stained. Twenty-seven enzymes were tested; 12 were expressed in H. influenzae. Six enzymes were selected for subsequent study: malate dehydrogenase (MDH), phenylalanylleucine peptidase (PE2), 6-phosphogluconate dehydrogenase (6PG), adenylate kinase (AK), glucose 6-phosphate dehydrogenase (G6P), and phosphoglucose isomerase (PGI). They were polymorphic and occurred in all isolates. Six electromorphs were found for PE2, G6P, and PGI, five for MDH, four for 6PG, and three for AK. PE2, G6P, and PGI contributed most of the ET resolution (48 of 49 ETs). Multilocus enzyme electrophoresis showed several advantages over previous typing techniques. An ET could be assigned to both typable and nontypable (NT) isolates. The technique was powerful in resolving differences among isolates. The 94 isolates comprised 49 ETs, five biotypes, and six capsular types and NT isolates. Strains known to be related expressed the same ET, e.g., RAB b+ and b-, ET12; Ma a+ and a-, ET1. ET variability among type b isolates was low; 26 of 28 clinical isolates expressed ET14; 2 of 28 expressed ET13 and ET15, differing from ET14 by one electromorph each. In contrast, the 47 NT isolates comprised 38 different ETs. No ETs were shared between non-type b capsulated strains and type b or NT strains. Interestingly, five NT isolates expressed the same ET as type b strains. (iv) Strains of the same capsular type but different biotypes expressed different ETs. ET determinations will thus be useful in studying the epidemiology and evolution of H. influenzae.